Su Haochen, Yuan Yue, Tang Jiatong, Zhang Yixuan, Wu Hao, Zhang Yin, Liang Jiawei, Wang Lei, Zou Xiaoping, Huang Shuling, Zhang Shu, Lv Ying
Department of Gastroenterology, Nanjing Drum Tower Hospital Clinical College of Nanjing Medical University, No. 321 Zhongshan Road, Nanjing, Jiangsu 210008, PR China; Institute of Pancreatology, Nanjing University, Nanjing, Jiangsu 210008, PR China.
Department of Gastroenterology, Nanjing Drum Tower Hospital Clinical College of Nanjing Medical University, No. 321 Zhongshan Road, Nanjing, Jiangsu 210008, PR China; Department of Gastroenterology, The Third People's Hospital of Yancheng, Yancheng, Jiangsu 224000, PR China.
Transl Oncol. 2023 Oct;36:101743. doi: 10.1016/j.tranon.2023.101743. Epub 2023 Jul 28.
Chemoresistance is a common event after cancer chemotherapy, including gastric cancer (GC). Cisplatin has been reported to induce the DNA damage response (DDR), thus leading to chemoresistance. VE-821, a specific inhibitor of ATR, has been proven to suppress a variety of solid malignancies effectively. Our study aimed to explore the effect of VE-821 on enhancing the chemical sensitivity to cisplatin and clarify the potential molecular mechanisms.
Cell viability and apoptosis of MKN-45 and AGS were measured by CCK8 and flow cytometry assay respectively. Western blotting was used to detect the expression of target proteins. TCGA database was used to analyze the correlation between the ATR expression with the prognosis of GC patients. The viability of GC organoids was detected by Cell Titer Glo (CTG) through luminescence.
Cisplatin inhibited the proliferation and induced apoptosis of GC cells with a relatively high IC50 value, and increased the phosphorylation levels of ATR-CHK1 and H2AX. VE-821 achieved the same effects but by downregulating the phosphorylation levels of the ATR-CHK1 pathway. Besides, higher ATR expression in GC tissues was positively correlated with higher pathological stage in GC patients. Interestingly, ATR inhibition reversed cisplatin-induced STAT3 activation and enhanced H2AX levels. Moreover, VE-821 significantly sensitized GC cells to cisplatin, and these two drugs had synergistic effects in GC cell lines, organoids, and in vivo.
Our results suggested VE-821 sensitized GC cells to cisplatin via reversing DDR activation. And VE-821 treatment may be a promising therapeutic strategy for GC patients with cisplatin resistance.
化疗耐药是癌症化疗（包括胃癌（GC））后的常见现象。据报道，顺铂可诱导DNA损伤反应（DDR），从而导致化疗耐药。VE-821是一种特异性的ATR抑制剂，已被证明能有效抑制多种实体恶性肿瘤。我们的研究旨在探讨VE-821对增强顺铂化学敏感性的作用，并阐明其潜在的分子机制。
分别采用CCK8法和流式细胞术检测MKN-45和AGS细胞的活力和凋亡情况。采用蛋白质免疫印迹法检测靶蛋白的表达。利用TCGA数据库分析ATR表达与GC患者预后的相关性。通过Cell Titer Glo（CTG）发光法检测GC类器官的活力。
顺铂以相对较高的IC50值抑制GC细胞增殖并诱导其凋亡，并增加ATR-CHK1和H2AX的磷酸化水平。VE-821通过下调ATR-CHK1通路的磷酸化水平达到相同效果。此外，GC组织中较高的ATR表达与GC患者较高的病理分期呈正相关。有趣的是，抑制ATR可逆转顺铂诱导的STAT3激活并增强H2AX水平。此外，VE-821显著增强了GC细胞对顺铂的敏感性，这两种药物在GC细胞系、类器官和体内均具有协同作用。
我们的结果表明，VE-821通过逆转DDR激活使GC细胞对顺铂敏感。VE-821治疗可能是顺铂耐药GC患者的一种有前景的治疗策略。
